Difference between revisions of "Part:BBa K1149008"

Revision as of 23:06, 2 October 2013

Proteinase K (extracellular)

Encodes a peptidase from proteinase K family, breaks down polylactic acid. See here for construct with secretion tag.

Characterisation

Our Proteinase K (with PelB secretion tag) construct contains sfGFP within an operon and therefore fluorescence can be utilised to determine if expression is being induced by addition of Xylose.

Proteinase K with pelB secretion tag growth assay. As with Proteinase K without pelB, there is increased growth compared to uninduced for BBa_K1149008. However this is not significant as a two-tailed t-test gave p = 0.4442. Growth was at 37°C with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.

Proteinase K with pelB secretion tag induction assay. Induction of BBa_K1149008 gives a very strong induction response, this is significant as a two-tailed t-test gave a value of p = 0.0104 < 0.05. Thus the induction does have a significant effect on fluorescence for Proteinase K with pelB. Growth was at 37°C with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.

Conclusion: There is no growth inhibition caused by induction, however induction is significantly induced by the Xylose promoter.

References:

Larsen AN, Moe E, Helland R, Gjellesvik DR, Willassen NP.Characterization of a recombinantly expressed proteinase K-like enzyme from a psychrotrophic Serratia sp. FEBS J. 2006 Jan;273(1):47-60.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 125
Illegal NheI site found at 1009
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 356
Illegal BamHI site found at 65
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 210
Illegal AgeI site found at 1531
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1149008 short</partinfo>
-Encodes a peptidase from proteinase K family, breaks down polylactic acid.
+Encodes a peptidase from proteinase K family, breaks down polylactic acid. See [https://parts.igem.org/Part:BBa_K1149007 here] for construct with secretion tag.
+<h2>Characterisation</h2>
+Our Proteinase K (with PelB secretion tag) construct contains sfGFP within an operon and therefore fluorescence can be utilised to determine if expression is being induced by addition of Xylose.
-reference:
+https://static.igem.org/mediawiki/2013/thumb/a/ab/PK_pelB.png/450px-PK_pelB.png
+'''Proteinase K with pelB secretion tag growth assay.''' As with Proteinase K without pelB, there is increased growth compared to uninduced for BBa_K1149008. However this is not significant as a two-tailed t-test gave p = 0.4442. Growth was at 37°C with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.
+https://static.igem.org/mediawiki/2013/thumb/5/54/PK_pelB_fluor.png/450px-PK_pelB_fluor.png
+'''Proteinase K with pelB secretion tag induction assay.''' Induction of BBa_K1149008 gives a very strong induction response, this is significant as a two-tailed t-test gave a value of p = 0.0104 < 0.05. Thus the induction does have a significant effect on fluorescence for Proteinase K with pelB. Growth was at 37°C with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.
+<b>Conclusion: There is no growth inhibition caused by induction, however induction is significantly induced by the Xylose promoter. </b>
+<h2>References: </h2>
 Larsen AN, Moe E, Helland R, Gjellesvik DR, Willassen NP.Characterization of a recombinantly expressed proteinase K-like enzyme from a psychrotrophic Serratia sp. FEBS J. 2006 Jan;273(1):47-60.